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• W2022105035 startingPage "56" @default.
• W2022105035 abstract "Alcohol dehydrogenases (ADH) are a class of oxidoreductases that catalyse the reversible oxidation of ethanol to acetaldehyde. In the human parasite Trypanosoma cruzi the TcADH gene was identified through microarray analysis as having reduced transcription in an invitro induced benznidazole (BZ)-resistant population. In the present study, we have extended these results by characterizing the TcADH gene from 11 strains of T. cruzi that were either susceptible or naturally resistant to benznidazole and nifurtimox or had in vivo selected or in vitro induced resistance to BZ. Sequence comparisons showed that TcADH was more similar to prokaryotic ADHs than to orthologs identified Leishmania spp. Immunolocalisation using confocal microscopy revealed that TcADH is present in the kinetoplast region and along the parasite body, consistent with the mitochondrial localization predicted by sequence analysis. Northern blots showed a 1.9 kb transcript with similar signal intensity in all T. cruzi samples analysed, except for the in vitro selected resistant population, where transcript levels were 2-fold lower. These findings were confirmed by quantitative real-time PCR. In Western blot analysis, anti-TcADH polyclonal antisera recognised a 42 kDa protein in all T. cruzi strains tested. The level of expression of this polypeptide was approximately 2-fold lower in the in vitro induced benznidazole-resistant strain, than in the susceptible parental strain. The chromosomal location of the TcADH gene was variable, but was not associated with the zymodeme or with the drug resistance phenotype. The data presented here show that the TcADH enzyme has a decreased level of expression in the in vitro induced BZ-resistant T. cruzi population, a situation that has not been observed in the in vivo selected BZ-resistant and naturally resistant strains." @default.
• W2022105035 created "2016-06-24" @default.
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• W2022105035 date "2009-07-01" @default.
• W2022105035 modified "2023-09-30" @default.
• W2022105035 title "Characterization of a gene encoding alcohol dehydrogenase in benznidazole-susceptible and -resistant populations of Trypanosoma cruzi" @default.
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• W2022105035 doi "https://doi.org/10.1016/j.actatropica.2009.02.007" @default.
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